Expanding the Caenorhabditis elegans auxin-inducible degron system toolkit with internal expression and degradation controls and improved modular constructs for CRISPR/Cas9-mediated genome editing Institutional Repository Document uri icon


  • AbstractThe auxin-inducible degron (AID) system has emerged as a powerful tool to conditionally deplete proteins in a range of organisms and cell-types. Here, we describe a toolkit to augment the use of the AID system inCaenorhabditis elegans. We have generated a set of single-copy, tissue-specific (germline, intestine, neuron, muscle, hypodermis, seam cell, anchor cell) and pan-somaticTIR1-expressing strains carrying an equimolar co-expressed blue fluorescent reporter to enable use of both red and green channels in experiments. We have also constructed a set of plasmids to generate fluorescent protein::AID fusions through CRISPR/Cas9-mediated genome editing. These templates can be produced through frequently used cloning systems (Gibson assembly or SapTrap) or through ribonucleoprotein complex-mediated insertion of PCR-derived, linear repair templates. We have generated a set of sgRNA plasmids carrying modifications shown to boost editing efficiency, targeting standardized transgene insertion sites on chromosomes I and II. Together these reagents should complement existingTIR1strains and facilitate rapid and high-throughput fluorescent protein::AID* tagging of factors of interest. This battery of new TIR1-expressing strains and modular, efficient cloning vectors serves as a platform for facile assembly of CRISPR/Cas9 repair templates for conditional protein depletion.

altmetric score

  • 45.584

author list (cited authors)

  • Ashley, G., Duong, T., Levenson, M. T., Martinez, M., Hibshman, J. D., Saeger, H. N., ... Ward, J. D.

citation count

  • 8

complete list of authors

  • Ashley, Guinevere||Duong, Tam||Levenson, Max T||Martinez, Michael AQ||Hibshman, Jonathan D||Saeger, Hannah N||Doonan, Ryan||Palmisano, Nicholas J||Martinez-Mendez, Raquel||Davidson, Brittany||Zhang, Wan||Ragle, James Matthew||Medwig-Kinney, Taylor N||Sirota, Sydney S||Goldstein, Bob||Matus, David Q||Dickinson, Daniel J||Reiner, David J||Ward, Jordan D

Book Title

  • bioRxiv

publication date

  • May 2020